The Determination of Naturally Produced Penicillins and Their Biosynthetic Precursors Using Pre-Column Derivatisation with Dansylaziridine

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Abstract
A novel method of pre-column derivatisation has been developed for the trace-level HPLC determination of penicillins and their biosynthetic intermediates in complex biological samples. The derivatisation is reliant on the reaction between dansylaziridine and a thiolate anion produced by alkaline degradation of the penicillin ring systems. This reaction results in the formation of stable fluorescent products, which may be detected spectrofluorimetrically. Optimisation of both the derivatisation reaction and the chromatographic conditions has been investigated. Using this technique we have been able to simultaneously measure both intraand extra-cellular levels of penicillins and their biosynthetic intermediates produced in fermentations at concentrations below 1 μg/ml.